The impact of the implementation of the Dutch combined Meeting Centres Support Programme for family caregivers of people with dementia in Italy, Poland and UK.
Objectives: The MEETINGDEM research project aimed to implement the combined Dutch Meeting Centre Support Programme (MCSP) for community-dwelling people with dementia and caregivers within Italy, Poland and UK and to assess whether comparable benefits were found in these countries as in the Netherlands.Method: Nine pilot Meeting Centres (MCs) participated (Italy-5, Poland-2, UK-2). Effectiveness of MCSP was compared to usual care (UC) on caregiver outcomes measuring competence (SSCQ), mental health (GHQ-12), emotional distress (NPI-Q) and loneliness (UCLA) analysed by ANCOVAs in a 6-month pre-test/post-test controlled trial. Interviews using standardised measures were completed with caregivers.Results: Pre/post data were collected for 93 caregivers receiving MCSP and 74 receiving UC. No statistically significant differences on the outcome measures were found overall. At a country level MC caregivers in Italy showed significant better general mental health (p = 0.04, d = 0.55) and less caregiver distress (p = 0.02, d = 0.62) at post-test than the UC group. Caregiver satisfaction was rated on a sample at 3 months (n = 81) and 6 months (n = 84). The majority of caregivers reported feeling less burdened and more supported by participating in MCSP.Conclusion: The moderate positive effect on sense of competence and the greater mental health benefit for lonely caregivers using the MCSP compared to UC as found in the original Dutch studies were not replicated. However, subject to study limitations, caregivers in Italy using MCSP benefitted more regarding their mental health and emotional distress than caregivers using UC. Further evaluation of the benefits of MCSP within these countries in larger study samples is recommended.


Mental disorders affect older persons in Sweden--a register-based study.
The study aimed to estimate the prevalence of mental disorders based on pharmaceutical use among the old (age ≥65) in Sweden for the years 2006-2008. Data on the mental health of older persons were approximated on the basis of recommended prescriptions for pharmaceuticals, gathered from the Swedish Register on Prescribed Pharmaceuticals (SRPP). Each disorder (ICD-10, F20-F42, and F60-F61) was analyzed to identify associated recommended pharmaceuticals. Anatomical Therapeutic Chemical Classification codes were applied. The data covered 188,024 individuals who received 2,013,079 prescriptions for pharmaceuticals for mental disorders during a 3-year period. Persons with pharmaceuticals for dementia disorders were excluded from the calculations of the prevalence of mental disorders. The prevalence of mental disorders among the old in Sweden, measured on the basis of pharmaceutical use, was 6.6% in 2006, 2007, and 2008, respectively. Men constituted one-third of cases and women two-thirds. Prevalence was lowest in the age group 65-69 and increased subsequently with age. This fundamental register-based study included a great number of older persons and shows that mental disorders affect every fifteenth older person in Sweden. The prevalence of mental disorders increases with increasing age. The results highlight the extent of mental disorders among older persons, which is important to know when planning care for these patients. This study, by investigating a large population, provides a solid basis for general planning as well as for future mental disorder research.


Gene therapy in glaucoma-part I: Basic mechanisms and molecular genetics.
Glaucoma is the second most common cause of blindness in the world as determined by the World Health Organization (WHO). Glaucoma diagnosis, identification of people at risk, initiation of treatment and timing of surgical intervention remains a problem. Despite new and improving diagnostic and therapeutic options for glaucoma, blindness from glaucoma remains a major public health problem. The role of heredity in ocular disease is attracting greater attention as knowledge and recent advances of Human Genome Project and the HapMap Project have made genetic analysis of many human disorders possible.Glaucoma offers a variety of potential targets for gene therapy. All risk factors for glaucoma and their underlying causes are potentially susceptible to modulation by gene transfer. The discovery of genes responsible for glaucoma has led to the development of new methods of Deoxyribonucleic acid (DNA)-based diagnosis and treatment. As genetic defects responsible for glaucoma are identified and the biochemical mechanisms underlying the disease are recognized, new methods of therapy can be developed. It is of utmost importance for ophthalmologists and glaucoma specialists to be familiar with and understand the basic molecular mechanisms, genes responsible for glaucoma and the ways of genetic treatment. METHOD OF LITERATURE SEARCH: The literature was searched on the Medline database using the Pubmed interface.


Transcutaneous bilirubinometry in the newborn infant: state of the art.
Hyperbilirubinemia in the newborn infant continues to challenge physicians. Clinical evaluation and treatment have evolved well-established principles over the past decade. This review examines neonatal bilirubin metabolism and focuses on a recently developed clinical diagnostic tool, the transcutaneous bilirubinometer. In spite of some limitations, the transcutaneous bilirubinometer can be best applied as a screening tool to identify healthy full-term infants who require serum bilirubin determination. With proper application, this device can eliminate most invasive diagnostic testing. Optimal use of the instrument requires the relationship between the serum bilirubin concentration and the transcutaneous bilirubinometer index to be determined for each device, institution, and population.


Variable timing of synaptic transmission in cerebellar unipolar brush cells.
The cerebellum ensures the smooth execution of movements, a task that requires accurate neural signaling on multiple time scales. Computational models of cerebellar timing mechanisms have suggested that temporal information in cerebellum-dependent behavioral tasks is in part computed locally in the cerebellar cortex. These models rely on the local generation of delayed signals spanning hundreds of milliseconds, yet the underlying neural mechanism remains elusive. Here we show that a granular layer interneuron, called the unipolar brush cell, is well suited to represent time intervals in a robust way in the cerebellar cortex. Unipolar brush cells exhibited delayed increases in excitatory synaptic input in response to presynaptic stimulation in mouse cerebellar slices. Depending on the frequency of stimulation, delays extended from zero up to hundreds of milliseconds. Such controllable protraction of delayed currents was the result of an unusual mode of synaptic integration, which was well described by a model of steady-state AMPA receptor activation. This functionality extends the capabilities of the cerebellum for adaptive control of behavior by facilitating appropriate output in a broad temporal window.


Climate change and temperature-dependent sex determination in reptiles.
Despite increasing concern over the possible impact of global temperature change, there is little empirical evidence of direct temperature effects on biotic interactions in natural systems. Clear assessment of the ecological and evolutionary impact of changing climatic temperature requires a natural system in which populations exhibit a direct unambiguous fitness response to thermal fluctuation. I monitored nests of a population of painted turtles (Chrysemys picta) with temperature-dependent sex determination to investigate the causal relationship between local climatic variation in temperature and offspring sex ratio. Consistent with theoretical predictions, annual offspring sex ratio was highly correlated with mean July air temperature, validating concerns about the effect of climate change on population demography. This correlation implies that even modest increases in mean temperature (< 2 degrees C) may drastically skew the sex ratio. Statistical evaluation of the variance in climate change indicates that an increase in mean temperature of 4 degrees C would effectively eliminate production of male offspring. Quantitative genetic analyses and behavioral data suggest that populations with temperature-dependent sex determination may be unable to evolve rapidly enough to counteract the negative fitness consequences of rapid global temperature change. Populations of species with temperature-dependent sex determination may serve as ideal indicators of the biological impact of global temperature change.


Mannan-based conjugates as a multimodal imaging platform for lymph nodes.
We show that mannan-based conjugates possess exceptional features for multimodal imaging because of their biocompatibility, biodegradability and self-targeting properties. Two new mannan conjugates, containing a gadolinium complex and a fluorescent probe, one based only on polysaccharide and the other one comprising polysaccharide with poly(2-methyl-2-oxazoline) grafts, were prepared and simultaneously visualized in vitro and in vivo by magnetic resonance and fluorescence imaging. The synthesis of these mannan-based complexes was based on alkylation with allyl bromide or grafting with poly(2-methyl-2-oxazoline) chains, followed by a thiol-ene click reaction with cysteamine to introduce primary amino groups into their structure. Finally, the obtained conjugates were functionalized with contrast labels using the corresponding N-hydroxysuccinimide esters. When used to detect lymph nodes, the polymers showed better imaging properties than a commercially available contrast agent.


Overexpression and purification of recombinant eRF1 proteins of rabbit and Tetrahymena thermophila.
The polypeptide release factor (eRF1) gene was cloned from rabbit and its overexpression and purification system was established in parallel with that of the eRF1 gene of Tetrahymena thermophila that has been cloned recently in this laboratory. The rabbit eRF1 (Ra-eRF1) is composed of 437 amino acids and is completely identical to human eRF1 though 3% distinct in the nucleotide sequence. This is in sharp contrast to Tetrahymena eRF1 (Tt-eRF1) that is only 57% identical to human eRF1. The recombinant Ra-eRF1 was marked with a histidine tag, overexpressed, and purified to homogeneity by two-step chromatography using Ni-NTA-agarose and Mono Q columns. In contrast to Ra-eRF1, Tt-eRF1 formed aggregates upon overexpression in Escherichia coli, hence it was purified under denaturing conditions, and used to raise rabbit antibody. The resulting anti-Tt-eRF1 antibody proved useful for examining conditions for soluble Tt-eRF1 in test cells. Finally, a soluble Tt-eRF1 fraction was purified from Saccharomyces cerevisiae transformed with the Tt-eRF1 expression plasmid by three steps of affinity and anion exchange chromatography. The cloned Ra-eRF1 gene complemented a temperature-sensitive allele in the eRF1 gene, sup45 (ts), of S. cerevisiae, though the complementation activity was significantly impaired by the histidine tag, whereas Tt-eRF1 failed to complement the sup45 (ts) allele.


Golgins and GTPases, giving identity and structure to the Golgi apparatus.
In this review we will focus on the recent advances in how coiled-coil proteins of the golgin family give identity and structure to the Golgi apparatus in animal cells. A number of recent studies reveal a common theme for the targeting of golgins containing the ARL-binding GRIP domain, and the related ARF-binding GRAB domain. Similarly, other golgins such as the vesicle tethering factor p115 and Bicaudal-D are targeted by the Rab GTPases, Rab1 and Rab6, respectively. Together golgins and their regulatory GTPases form a complex network, commonly known as the Golgi matrix, which organizes Golgi membranes and regulates membrane trafficking.


Application of latent growth and growth mixture modeling to identify and characterize differential responders to treatment for COPD.
To explore the utility of applying growth mixture models (GMMs) in secondary analyses of clinical trials to identify sources of variability in data reported by patients with COPD. Analyses were performed on data from two 6-month clinical trials comparing indacaterol and open-label tiotropium or blinded salmeterol and the first six months of a 12-month trial comparing indacaterol and blinded formoterol. Latent growth model (LGM) analyses were conducted to explore the response of the SGRQ Symptoms score from baseline to six months and GMM analyses were evaluated as a method to identify latent classes of differential responders. Variability in SGRQ Symptom scores was found suggesting subsets of patients with differential response to treatment. GMM analyses found subsets of non-responders in all trials. When the responders were analyzed separately from non-responders, there were increased treatment effects (e.g., symptoms score improvement over six months for whole groups: indacaterol=8-12 units, tiotropium=7 units, salmeterol=9 units, formoterol=11 units. Responder subgroup improvement: indacaterol=9-21 units, tiotropium=7 units, salmeterol=10 units, formoterol=20 units). Responders had significantly different baseline SGRQ Symptom scores, smoking history, age, and mMRC dyspnea scores than non-responders. Patients with COPD represent a heterogeneous population in terms of their reporting of symptoms and response to treatment. GMM analyses are able to identify sub-groups of responders and non-responders. Application of this methodology could be of value on other endpoints in COPD and in other disease areas.


Effect of metformin therapy on circulating amino acids in a randomized trial: the CAMERA study.
To investigate whether metformin therapy alters circulating aromatic and branched-chain amino acid concentrations, increased levels amino acid concentrations, increased levels of which have been found to predict Type 2 diabetes. In the Carotid Atherosclerosis: Metformin for Insulin Resistance (CAMERA) study (NCT00723307), 173 individuals without Type 2 diabetes, but with coronary disease, were randomized to metformin (n=86) or placebo (n=87) for 18 months. Plasma samples, taken every 6 months, were analysed using quantitative nuclear magnetic resonance spectroscopy. Ten metabolites consisting of eight amino acids [three branched-chain (isoleucine, leucine, valine), three aromatic (tyrosine, phenylalanine, histidine) and two other amino acids (alanine, glutamine)], lactate and pyruvate were quantified and analysed using repeated-measures models. On-treatment analyses were conducted to investigate whether amino acid changes were dependent on changes in weight, fat mass or insulin resistance estimated using homeostasis model assessment (HOMA-IR). Tyrosine decreased [-6.1 μmol/l (95% CI -8.5, -3.7); P<0.0001], while alanine [42 umol/l (95% CI 25, 59); P<0.0001] increased in the metformin-treated group compared with the placebo-treated group. Decreases in phenylalanine [-2.0 μmol/l (95% CI -3.6, -0.3); P=0.018] and increases in histidine [2.3 μmol/l (95% CI 0.1, 4.6); P=0.045] were also observed in the metformin group, although these changes were less statistically robust. Changes in these four amino acids were not accounted for by changes in weight, fat mass or HOMA-IR values. Levels of branched-chain amino acids, glutamine, pyruvate and lactate were not altered by metformin therapy. Metformin therapy results in a sustained and specific pattern of changes in aromatic amino acid and alanine concentrations. These changes are independent of any effects on weight and insulin sensitivity. Any causal link to metformin's unexplained cardiometabolic benefit requires further study.


Cytokine Profiles during Invasive Nontyphoidal Salmonella Disease Predict Outcome in African Children.
Nontyphoidal Salmonella is a leading cause of sepsis in African children. Cytokine responses are central to the pathophysiology of sepsis and predict sepsis outcome in other settings. In this study, we investigated cytokine responses to invasive nontyphoidal Salmonella (iNTS) disease in Malawian children. We determined serum concentrations of 48 cytokines with multiplexed immunoassays in Malawian children during acute iNTS disease (n = 111) and in convalescence (n = 77). Principal component analysis and logistic regression were used to identify cytokine signatures of acute iNTS disease. We further investigated whether these responses are altered by HIV coinfection or severe malnutrition and whether cytokine responses predict inpatient mortality. Cytokine changes in acute iNTS disease were associated with two distinct cytokine signatures. The first is characterized by increased concentrations of mediators known to be associated with macrophage function, and the second is characterized by raised pro- and anti-inflammatory cytokines typical of responses reported in sepsis secondary to diverse pathogens. These cytokine responses were largely unaltered by either severe malnutrition or HIV coinfection. Children with fatal disease had a distinctive cytokine profile, characterized by raised mediators known to be associated with neutrophil function. In conclusion, cytokine responses to acute iNTS infection in Malawian children are reflective of both the cytokine storm typical of sepsis secondary to diverse pathogens and the intramacrophage replicative niche of NTS. The cytokine profile predictive of fatal disease supports a key role of neutrophils in the pathogenesis of NTS sepsis.


Prevalence of chronic kidney disease in Asia: a systematic review and analysis.
The burden of chronic kidney disease (CKD) is growing rapidly around the world. However, there is limited information on the overall regional prevalence of CKD, as well as the variations in national prevalence within Asia. We aimed to consolidate available data and quantify estimates of the CKD burden in this region. We systematically searched MEDLINE, Embase and Google Scholar for observational studies and contacted national experts to estimate CKD prevalence in countries of Asia (Eastern, Southern and South Eastern Asia). CKD was defined as estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m2 or the presence of proteinuria. For countries without reported data, we estimated CKD prevalence using agglomerative average-linkage hierarchical clustering, based on country-level risk factors and random effects meta-analysis within clusters. Published CKD prevalence data were obtained for 16 countries (of the 26 countries in the region) and estimates were made for 10 countries. There was substantial variation in overall and advanced (eGFR <30 mL/min/1.73 m2) CKD prevalence (range: 7.0%-34.3% and 0.1%-17.0%, respectively). Up to an estimated 434.3 million (95% CI 350.2 to 519.7) adults have CKD in Asia, including up to 65.6 million (95% CI 42.2 to 94.9) who have advanced CKD. The greatest number of adults living with CKD were in China (up to 159.8 million, 95% CI 146.6 to 174.1) and India (up to 140.2 million, 95% CI 110.7 to 169.7), collectively having 69.1% of the total number of adults with CKD in the region. The large number of people with CKD, and the substantial number with advanced CKD, show the need for urgent collaborative action in Asia to prevent and manage CKD and its complications.


Associations between work environment, health status and sick leave among pregnant employees.
Aims: To study the associations between and timing of psychosocial and physical work factors and health status on sick leave among Danish pregnant employees. Methods: A total of 910 pregnant women completed a questionnaire in gestational weeks 12 (baseline) and 27 (follow-up). Information about psychosocial and physical work factors and health status was obtained at baseline. Associations with sick leave ⩾14 days were estimated using logistic regression. Further, the impact of timing and duration of exposure on sick leave were examined. Results: A total of 133 women (14.6%) reported ⩾14 days of sick leave at follow-up (27 weeks of gestation). Work-related risk factors for sick leave were high work pace, low influence, low recognition, low job satisfaction, conflict in work-family balance, standing/walking, heavy lifting, and shift work/night shift. Health-related risk factors were burnout, stress, possibility of depression, low work ability, previous sick leave, and poor self-rated health. Being exposed to work-related risk factors during the first 27 weeks of pregnancy or at follow-up increased the risk of sick leave compared with those not exposed at any time or only exposed at baseline. Poor health status increased the risk if women were exposed in the first 27 weeks of pregnancy; however, high possibility of depression was also a risk factor when experienced in early pregnancy. Conclusions: Psychosocial and physical work-related risk factors and poor health status were associated with more sick leave in pregnant employees. Early adjustment of work-related risk factors at the workplace is needed to reduce sick leave.


Modulating inflammation in a cutaneous chronic wound model by IL-10 released from collagen-silica nanocomposites via gene delivery.
Cutaneous chronic wounds remain a major clinical challenge which requires the development of novel wound dressings. Previously, we showed that collagen-silica nanocomposites consisting of polyethyleneimine (PEI)-DNA complexes associated with silica nanoparticles (SiNP), collagen hydrogel and 3T3 fibroblasts, can work as a local "cell factory". Indeed, the "in-gel" transfection leads to a sustained production and release of biomolecules. Herein, we further explored the possibility for nanocomposites to deliver interleukin-10 (IL-10), a potent anti-inflammatory cytokine, which favors tissue repair. Its anti-inflammatory effect was evaluated in an in vitro inflammation model carried out by LPS (lipopolysaccharide) activation of macrophages embedded in collagen gel. The IL-10 synthesis from nanocomposites was detected over one week in the range of 200-400 pg mL-1 and reached a maximum at day 5 without any observed cytotoxic effects. PEI10-SiNP outperformed free PEI10 and PEI25-SiNP, implying that the introduction of SiNP improved the transfection efficiency of low Mw of PEI. In addition, the structure and mechanical properties of collagen-silica nanocomposites were stable over one week. Subsequently, the ability of nanocomposites to modulate inflammation was tested in a 3D model of inflammation. The decrease of TNF-α and IL-1β gene expression by 20-80% indicated successful inhibition of inflammation by IL-10 released from nanocomposites. Taken together, the nanocomposites are capable of producing effective doses of IL-10 which inhibit the synthesis of pro-inflammatory cytokines and favor the expression of wound healing cytokines. Therefore, the as-constructed 3D gene delivery system represents a promising strategy for the controlled release of therapeutic biomolecules favoring cutaneous wound healing.


Dynamic imaging of the immune system: progress, pitfalls and promise.
Both innate and adaptive immunity are dependent on the migratory capacity of myeloid and lymphoid cells. Effector cells of the innate immune system rapidly enter infected tissues, whereas sentinel dendritic cells in these sites mobilize and transit to lymph nodes. In these and other secondary lymphoid tissues, interactions among various cell types promote adaptive humoral and cell-mediated immune responses. Recent advances in light microscopy have allowed direct visualization of these events in living animals and tissue explants, which allows a new appreciation of the dynamics of immune-cell behaviour. In this article, we review the basic techniques and the tools used for in situ imaging, as well as the limitations and potential artefacts of these methods.


gamma-Tocotrienol prevents oxidative stress-induced telomere shortening in human fibroblasts derived from different aged individuals.
The effects of palm gamma-tocotrienol (GGT) on oxidative stress-induced cellular ageing was investigated in normal human skin fibroblast cell lines derived from different age groups; young (21-year-old, YF), middle (40-year-old, MF) and old (68-year-old, OF). Fibroblast cells were treated with gamma-tocotrienol for 24 hours before or after incubation with IC50 dose of H2O2 for 2 hours. Changes in cell viability, telomere length and telomerase activity were assessed using the MTS assay (Promega, USA), Southern blot analysis and telomere repeat amplification protocol respectively. Results showed that treatment with different concentrations of gamma-tocotrienol increased fibroblasts viability with optimum dose of 80 microM for YF and 40 microM for both MF and OF. At higher concentrations, gamma-tocotrienol treatment caused marked decrease in cell viability with IC50 value of 200 microM (YF), 300 microM (MF) and 100 microM (OF). Exposure to H2O2 decreased cell viability in dose dependent manner, shortened telomere length and reduced telomerase activity in all age groups. The IC50 of H2O2 was found to be; YF (700 microM), MF (400 microM) and OF (100 microM). Results showed that viability increased significantly (p < 0.05) when cells were treated with 80 microM and 40 microM gamma-tocotrienol prior or after H2O2-induced oxidative stress in all age groups. In YF and OF, pretreatment with gamma-tocotrienol prevented shortening of telomere length and reduction in telomerase activity. In MF, telomerase activity increased while no changes in telomere length was observed. However, post-treatment of gamma-tocotrienol did not exert any significant effects on telomere length and telomerase activity. Thus, these data suggest that gamma-tocotrienol protects against oxidative stress-induced cellular ageing by modulating the telomere length possibly via telomerase.


Comparative investigation of fine bubble and macrobubble aeration on gas utility and biotransformation productivity.
The sufficient provision of oxygen is mandatory for enzymatic oxidations in aqueous solution, however, in process optimization this still is a bottleneck that cannot be overcome with the established methods of macrobubble aeration. Providing higher mass transfer performance through microbubble aerators, inefficient aeration can be overcome or improved. Investigating the mass transport performance in a model protein solution, the microbubble aeration results in higher kL a values related to the applied airstream in comparison with macrobubble aeration. Comparing the aerators at identical kL a of 160 and 60 1/h, the microbubble aeration is resulting in 25 and 44 times enhanced gas utility compared with aeration with macrobubbles. To prove the feasibility of microbubbles in biocatalysis, the productivity of a glucose oxidase catalyzed biotransformation is compared with macrobubble aeration as well as the gas-saving potential. In contrast to the expectation that the same productivities are achieved at identically applied kL a, microbubble aeration increased the gluconic acid productivity by 32% and resulted in 41.6 times higher oxygen utilization. The observed advantages of microbubble aeration are based on the large volume-specific interfacial area combined with a prolonged residence time, which results in a high mass transfer performance, less enzyme deactivation by foam formation, and reduced gas consumption. This makes microbubble aerators favorable for application in biocatalysis.


Applicability of Progress Testing in Veterinary Medical Education.
A substantial part of graduate education in veterinary medicine is spent in clinical practice. During the clinical experiential phase, it is difficult to monitor students' actual knowledge development: they build individual records of experiences based on the cases they have to deal with, while mainly focusing on knowledge that is of direct, clinical relevance to them. As a result, students' knowledge bases may differ to such a degree that a single test alone may not be able to provide an adequate reflection of progress made. In these circumstances, progress testing, which is a method of longitudinal assessment independent of the curricular structure, may offer a viable solution. The purpose of this study, therefore, was to determine the extent to which progress tests (PT) can be used to monitor progress in knowledge development at a graduate level in veterinary medical education. With a 6-month interval, we administered two tests to students based on the Maastricht Progress Test format that covered a large variety of veterinary topics. Consequently, we analyzed students' progress in knowledge development. Based on a substantive appraisal of the questions and analysis of the test results, we concluded that the tests met the measurement criteria. They appeared sensitive enough to gauge the progress made and were appreciated by the students. Hence, in spite of the differences within the whole graduate group, the PT format can be used to monitor students' knowledge development.


Effects of Dapagliflozin on Volume Status When Added to Renin-Angiotensin System Inhibitors.
Sodium glucose co-transporter 2 (SGLT2) inhibitors reduce the risk of heart and kidney failure in patients with type 2 diabetes, possibly due to diuretic effects. Previous non-placebo-controlled studies with SGLT2 inhibitors observed changes in volume markers in healthy individuals and in patients with type 2 diabetes with preserved kidney function. It is unclear whether patients with type 2 diabetes and signs of kidney damage show similar changes. Therefore, a post hoc analysis was performed on two randomized controlled trials (n = 69), assessing effects of dapagliflozin 10 mg/day when added to renin-angiotensin system inhibition in patients with type 2 diabetes and urinary albumin-to-creatinine ratio ≥30 mg/g. Blood and 24-h urine was collected at the start and the end of treatment periods lasting six and 12 weeks. Effects of dapagliflozin compared to placebo on various markers of volume status were determined. Fractional lithium excretion, a marker of proximal tubular sodium reabsorption, was assessed in 33 patients. Dapagliflozin increased urinary glucose excretion by 217.2 mmol/24 h (95% confidence interval (CI): from 155.7 to 278.7, p < 0.01) and urinary osmolality by 60.4 mOsmol/kg (from 30.0 to 90.9, p < 0.01), compared to placebo. Fractional lithium excretion increased by 19.6% (from 6.7 to 34.2; p < 0.01), suggesting inhibition of sodium reabsorption in the proximal tubule. Renin and copeptin increased by 46.9% (from 21.6 to 77.4, p < 0.01) and 33.0% (from 23.9 to 42.7, p < 0.01), respectively. Free water clearance (FWC) decreased by -885.3 mL/24 h (from -1156.2 to -614.3, p < 0.01). These changes in markers of volume status suggest that dapagliflozin exerts both osmotic and natriuretic diuretic effects in patients with type 2 diabetes and kidney damage, as reflected by increased urinary osmolality and fractional lithium excretion. As a result, compensating mechanisms are activated to retain sodium and water.


Contrast-enhanced Magnetic Resonance Imaging of Pelvic Bone Metastases at 3.0 T: Comparison Between 3-dimensional T1-weighted CAIPIRINHA-VIBE Sequence and 2-dimensional T1-weighted Turbo Spin-Echo Sequence.
This study aimed to compare 3-dimensional T1-weighted gradient-echo sequence (CAIPIRINHA-volumetric interpolated breath-hold examination [VIBE]) with 2-dimensional T1-weighted turbo spin-echo sequence for contrast-enhanced magnetic resonance imaging (MRI) of pelvic bone metastases at 3.0 T. Thirty-one contrast-enhanced MRIs of pelvic bone metastases were included. Two contrast-enhanced sequences were evaluated for the following parameters: overall image quality, sharpness of pelvic bone, iliac vessel clarity, artifact severity, and conspicuity and edge sharpness of the smallest metastases. Quantitative analysis was performed by calculating signal-to-noise ratio and contrast-to-noise ratio of the smallest metastases. Significant differences between the 2 sequences were assessed. CAIPIRINHA-VIBE had higher scores for overall image quality, pelvic bone sharpness, iliac vessel clarity, and edge sharpness of the metastatic lesions, and had less artifacts (all P < 0.05). There was no significant difference in conspicuity, signal-to-noise ratio, or contrast-to-noise ratio of the smallest metastases (P > 0.05). Our results suggest that CAIPIRINHA-VIBE may be superior to turbo spin-echo for contrast-enhanced MRI of pelvic bone metastases at 3.0 T.


Evaluation of the effects of repeated hand washing, sunlight, smoke and dirt on the persistence of deltamethrin on insecticide-treated nets.
Field studies were carried out in Iran to evaluate the effect of various factors (washing, sun, smoke, dust and dirt) on the residual insecticidal activity of PermaNet (a brand of long-lasting insecticidal net), and on nets conventionally treated with deltamethrin (K-O Tab), using bioassay tests. Thirty-two nets were washed five or 15 times, and eight nets were not washed at all. Nets were washed vigorously in cold tap water (17 degrees C, pH 8.9) with a detergent. Hand rubbing continued for 3min. After washing, some nets were exposed to dense smoke from a dung-hay fire for 3min and were also left exposed to the dusty wind between washes. One group of nets was exposed to the sunlight for the full 3-d interval between washes; another was exposed to sunlight for just 3h after each wash; two other groups were kept in the shade. There was a significantly greater loss of activity in nets exposed to the sun throughout the 3-d interval between washes: that is, for a total of 15 to 45 d. However, short sunlight exposure (maximum 3h between washes) during drying did not have any effect. We did not find any significant effect of exposure to dirt, dust and smoke after washing. It is concluded that the effect of sun is much smaller than that of washing, and that drying nets for a few hours in the sun is not harmful.


Positron Emission Tomography Imaging With [18F]flortaucipir and Postmortem Assessment of Alzheimer Disease Neuropathologic Changes.
Positron emission tomography (PET) may increase the diagnostic accuracy and confirm the underlying neuropathologic changes of Alzheimer disease (AD). To determine the accuracy of antemortem [18F]flortaucipir PET images for predicting the presence of AD-type tau pathology at autopsy. This diagnostic study (A16 primary cohort) was conducted from October 2015 to June 2018 at 28 study sites (27 in US sites and 1 in Australia). Individuals with a terminal illness who were older than 50 years and had a projected life expectancy of less than 6 months were enrolled. All participants underwent [18F]flortaucipir PET imaging, and scans were interpreted by 5 independent nuclear medicine physicians or radiologists. Supplemental autopsy [18F]flortaucipir images and pathological samples were also collected from 16 historically collected cases. A second study (FR01 validation study) was conducted from March 26 to April 26, 2019, in which 5 new readers assessed the original PET images for comparison to autopsy. [18F]flortaucipir PET images were visually assessed and compared with immunohistochemical tau pathology. An AD tau pattern of flortaucipir retention was assessed for correspondence with a postmortem B3-level (Braak stage V or VI) pathological pattern of tau accumulation and to the presence of amyloid-β plaques sufficient to meet the criteria for high levels of AD neuropathological change. Success was defined as having at least 3 of the 5 readers above the lower bounds of the 95% CI for both sensitivity and specificity of 50% or greater. A total of 156 patients were enrolled in the A16 study and underwent [18F]flortaucipir PET imaging. Of these, 73 died during the study, and valid autopsies were performed for 67 of these patients. Three autopsies were evaluated as test cases and removed from the primary cohort (n = 64). Of the 64 primary cohort patients, 34 (53%) were women and 62 (97%) were white; mean (SD) age was 82.5 (9.6) years; and 49 (77%) had dementia, 1 (2%) had mild cognitive impairment, and 14 (22%) had normal cognition. Prespecified success criteria were met for the A16 primary cohort. The flortaucipir PET scans predicted a B3 level of tau pathology, with sensitivity ranging from 92.3% (95% CI, 79.7%-97.3%) to 100.0% (95% CI, 91.0%-100.0%) and specificity ranging from 52.0% (95% CI, 33.5%-70.0%) to 92.0% (95% CI, 75.0%-97.8%). A high level of AD neuropathological change was predicted with sensitivity of 94.7% (95% CI, 82.7%-98.5%) to 100.0% (95% CI, 90.8%-100.0%) and specificity of 50.0% (95% CI, 32.1%-67.9%) to 92.3% (95% CI, 75.9%-97.9%). The FR01 validation study also met prespecified success criteria. Addition of the supplemental autopsy data set and 3 test cases, which comprised a total of 82 patients and autopsies for both the A16 and FR01 studies, resulted in improved specificity and comparable overall accuracy. Among the 156 enrolled participants, 14 (9%) experienced at least 1 treatment-emergent adverse event. This study's findings suggest that PET imaging with [18F]flortaucipir could be used to identify the density and distribution of AD-type tau pathology and the presence of high levels of AD neuropathological change, supporting a neuropathological diagnosis of AD.


Physical activity and lung cancer risk in the European Prospective Investigation into Cancer and Nutrition Cohort.
Research conducted predominantly in male populations on physical activity and lung cancer has yielded inconsistent results. We examined this relationship among 416,277 men and women from the European Prospective Investigation into Cancer and Nutrition (EPIC). Detailed information on recent recreational, household and occupational physical activity, smoking habits and diet was assessed at baseline between 1992 and 2000. Relative risks (RR) were estimated using Cox regression. During 6.3 years of follow-up we identified 607 men and 476 women with incident lung cancer. We did not observe an inverse association between recent occupational, recreational or household physical activity and lung cancer risk in either males or females. However, we found some reduction in lung cancer risk associated with sports in males (adjusted RR = 0.71; 95% confidence interval 0.50-0.98; highest tertile vs. inactive group), cycling (RR = 0.73; 0.54-0.99) in females and non-occupational vigorous physical activity. For occupational physical activity, lung cancer risk was increased for unemployed men (adjusted RR = 1.57; 1.20-2.05) and men with standing occupations (RR = 1.35; 1.02-1.79) compared with sitting professions. There was no evidence of heterogeneity of physical activity associations across countries, or across any of the considered cofactors. For some histologic subtypes suggestive sex-specific reductions, limited by subgroup sizes, were observed, especially with vigorous physical activity. In total, our study shows no consistent protective associations of physical activity with lung cancer risk. It can be assumed that the elevated risks found for occupational physical activity are not produced mechanistically by physical activity itself but rather reflect exposure to occupation-related lung cancer risk factors.


Design of diffractive singlets for monochromatic imaging.
The Seidel aberrations of a rotationally-symmetric diffractive lens with an arbitrary phase profile are presented. It is shown that by a proper choice of phase function and aperture stop position, third-order coma and astigmatism can be eliminated for any chosen conjugate ratio. Since a diffractive lens has an inherent zero value for the Petzval sum, the image plane is flat in both tangential and sagittal meridians. The substrate curvature of the lens may be chosen to introduce a prescribed amount of distortion to allow for use as a Fourier transform lens or a laser scan lens. Examples are given of lens performance in finite conjugate imaging and laser scanning, where the f - theta condition is satisfied.


Modulation of the Activities of Neuronal Ion Channels by Fatty Acid-Derived Pro-Resolvents.
Progress of inflammation depends on the balance between two biological mechanisms: pro-inflammatory and pro-resolving processes. Many extracellular and intracellular molecular components including cytokines, growth factors, steroids, neurotransmitters, and lipidergic mediators and their receptors contribute to the two processes, generated from cellular participants during inflammation. Fatty acid-derived mediators are crucial in directing the inflammatory phase and orchestrating heterogeneous reactions of participants such as inflamed cells, innate immune cells, vascular components, innervating neurons, etc. As well as activating specific types of receptor molecules, lipidergic mediators can actively control the functions of various ion channels via direct binding and/or signal transduction, thereby altering cellular functions. Lipid mediators can be divided into two classes based on which of the two processes they promote: pro-inflammatory, which includes prostaglandins and leukotrienes, and pro-resolving, which includes lipoxins, resolvins, and maresins. The research on the modulations of neuronal ion channels regarding the actions of the pro-inflammatory class has begun relatively earlier while the focus is currently expanding to cover the ion channel interaction with pro-resolvents. As a result, knowledge of inhibitory mechanisms by the pro-resolvents, historically seldom found for other known endogenous modulators or pro-inflammatory mediators, is accumulating particularly upon sensory neuronal cation channels. Diverse mechanistic explanations at molecular levels are being proposed and refined. Here we overviewed the interactions of lipidergic pro-resolvents with neuronal ion channels and outcomes from the interactions, focusing on transient receptor potential (TRP) ion channels. We also discuss unanswered hypotheses and perspectives regarding their interactions.


A large prospective study of SEP15 genetic variation, interaction with plasma selenium levels, and prostate cancer risk and survival.
The role of selenium in prostate cancer (PCa) risk remains controversial, but many epidemiologic studies suggest an inverse association with more aggressive disease. A recently discovered selenoprotein, SEP15, which is highly expressed in the prostate, may play a role either independently or by modifying the effects of selenium. We genotyped four common single-nucleotide polymorphisms capturing common variation (frequency >5%; R(2) > 0.8) within SEP15, as well as rs5859 in the 3' untranslated region, previously reported to reduce the efficiency of selenium incorporation into SEP15. We examined the association of these single-nucleotide polymorphisms with PCa risk and PCa-specific mortality, as well as their interactions with plasma selenium levels, in the Physicians' Health Study. In this nested case-control study (1,286 cases and 1,267 controls), SEP15 polymorphisms were not significantly associated with PCa risk. However, among the cases, three variants were significantly associated with PCa-specific mortality [rs479341 hazard ratio (HR), 1.94; 95% confidence interval (95% CI), 1.15-3.25; rs1407131 HR, 2.85; 95% CI, 1.45-5.59; rs561104 HR, 1.54; 95% CI, 1.12-2.11] with a recessive model. Additionally, rs561104 significantly modified the association of plasma selenium with PCa survival (P(interaction) = 0.02); an inverse relationship of high levels of selenium with PCa mortality was apparent only among those without the increased risk genotype. This study provides evidence that SEP15 genetic variation may influence PCa mortality. Additionally, the association of selenium with PCa mortality was modified by a variant, suggesting the possibility that some men with PCa may benefit more from selenium than others, depending on their genotype.


Depolarization and retardation of a birefringent slab.
Mueller matrices for normal transmission of light through a birefringent slab are formulated to analyze retardation and depolarization. A finite wave band, wedge slab, and microroughness may cause a spread in retardance, which in turn produces depolarization. The spectra of depolarization, cross-polarized transmittance, and retardance by rotating-analyzer ellipsometry are simulated for the quasi-monochromatic effect with a finite bandwidth. These spectra agree excellently with the measured spectra for a sapphire slab. The depolarization spectrum simulated for the wedge effect fits the measured spectrum in the long-wave region but is too small in the short-wave region. The depolarization simulated for incoherent multiple reflections demonstrates the oscillating structure, which is small compared with the measured depolarization. The finite bandwidth effect contributes more than the other effects to the measured depolarization of a sapphire slab.


Mantle oxidation state and its relationship to tectonic environment and fluid speciation.
The earth's mantle is degassed along mid-ocean ridges, while rehydration and possibly recarbonaton occurs at subduction zones. These processes and the speciation of C-H-O fluids in the mantle are related to the oxidation state of mantle peridotite. Peridotite xenoliths from continental localities exhibit an oxygen fugacity (fo(2)) range from -1.5 to +1.5 log units relative to the FMQ (fayalite-magnetite-quartz) buffer. The lowest values are from zones of continental extension. Highly oxidized xenoliths (fo(2) greater than FMQ) come from regions of recent or acive subduction (for example, Ichinomegata, Japan), are commonly amphibole-bearing, and show trace element and isotopic evidence of fluid-rock interaction. Peridotites from ocean ridges are reduced and have an averae fo(2) of about -0.9 log units relative to FMQ, virtually coincident with values obtained from mid-ocean ridge basalt (MORB) glasses. These data are further evidence of the genetic link between MORB liquids and residual peridotite and indicate that the asthenosphere, although reducing, has CO(2) and H(2)O as its major fluid species. Incorporation of oxidized material from subduction zones into the continental lithosphere produces xenoliths that have both asthenospheric and subduction signatures. Fluids in the lithosphere are also dominated by CO(2) and H(2)O, and native C is generally unstable. Although the occurrence of native C (diamond) in deep-seated garnetiferous xenoliths and kimberlites does not require reducing conditions, calculations indicate that high Fe(3+) contents are stabilized in the garnet structure and that fo(2) deareases with increasing depth.


All cause mortality and incidence of cancer in workers in bauxite mines and alumina refineries.
Bauxite is a reddish clay that is refined to produce alumina, which is then reduced to aluminium. There have been studies examining the health of workers in aluminium smelters, but not workers in bauxite mining and alumina refining. A cohort of employees of 1 large aluminium company since 1983 was assembled (n = 6,485, 5,828 men). Deaths and incident cancers to 2002 were ascertained by linkage to national and state cancer and death registries. SIRs and SMRs were calculated compared to national rates standardizing for calendar year, sex and 5-year age group. The mortality from all causes (SMR 0.68, 95% CI: 0.60-0.77), and from circulatory and respiratory diseases, all cancers combined and injury in the male cohort were lower than in the Australian male population and were similar across work groups and with duration of employment. The only significant increased mortality risk was from pleural mesothelioma. The incidence of all cancers combined was similar to the Australian rate. The cohort had a lower risk of incident lymphohaematopoietic cancer (SIR 0.50, 95% CI: 0.31-0.88) and a higher risk of melanoma (SIR 1.30, 95% CI: 1.00-1.69) although no dose-responses were seen. There was also an increased risk of mesothelioma (SIR 3.49, 95% CI: 1.82-6.71), which was associated with exposures outside the aluminium industry. This study is the first to examine cancer and mortality amongst workers in bauxite mines and alumina refineries and found little evidence for increased cancer incidence or mortality in these workers.


Melleolides induce rapid cell death in human primary monocytes and cancer cells.
The melleolides are structurally unique and bioactive natural products of the basidiomycete genus Armillaria. Here, we report on cytotoxic effects of melleolides from Armillaria mellea towards non-transformed human primary monocytes and human cancer cell lines, respectively. In contrast to staurosporine or pretubulysin that are less cytotoxic for monocytes, the cytotoxic potency of the active melleolides in primary monocytes is comparable to that in cancer cells. The onset of the cytotoxic effects of melleolides was rapid (within <1 h), as compared to the apoptosis inducer staurosporine, the protein biosynthesis inhibitor cycloheximide, and the DNA transcription inhibitor actinomycin D (>5 h, each). Side-by-side comparison with the detergent triton X-100 and staurosporine in microscopic and flow cytometric analysis studies as well as analysis of the viability of mitochondria exclude cell lysis and apoptosis as relevant or primary mechanisms. Our results rather point to necrotic features of cell death mediated by an as yet elusive but rapid mechanism.


A comparison between blood flow outcomes of tunneled external jugular and internal jugular hemodialysis catheters.
The right internal jugular (RIJ) vein is preferred for placement of long-term venous catheters (CVC) for hemodialysis (HD). Use of left IJ vein potentially depletes the access site for arteriovenous fistula (AVF) or arteriovenous graft (AVF) placement because of the high rates of central venous stenosis. The right external jugular (EJ) vein is a viable alternative to the LIJ. The purpose of this study is to compare blood flow outcomes in a series of percutaneously placed external jugular (EJ), LIJ and RIJ HD catheters. Using a prospectively collected database, we identified 46 hemodialysis patients who received a tunneled catheter during a 4-year period. Blood flow outcomes of RIJ, LIJ, and EJ catheters that were date-matched over the study period were compared. Using ANOVA, the blood flow outcomes of the 3 tunneled catheter techniques at 30-d and 90-d were compared. The 90-d blood flow outcomes of the 3 groups did not differ significantly. The 30-d blood flow was found to be 348.5± 56.62, 341± 22.42, and 365.7± 71.76 mL/min for RIJ, LIJ, and EJ respectively (P<.05). Using multiple regression analysis, no covariates (age, sex, race, diabetes) were found to be associated with blood flow outcomes at 30-d or 90-d. No identifiable factors were found to be associated with the difference in blood flow between 30-d and 90-d either. EJ blood flow outcomes at 30-d and 90-d were comparable to both LIJ and RIJ historic data. Further prospective investigation is required to define the role of EJ CVC placement as another potential long-term access modality.


Bottlenose dolphins can use learned vocal labels to address each other.
In animal communication research, vocal labeling refers to incidents in which an animal consistently uses a specific acoustic signal when presented with a specific object or class of objects. Labeling with learned signals is a foundation of human language but is notably rare in nonhuman communication systems. In natural animal systems, labeling often occurs with signals that are not influenced by learning, such as in alarm and food calling. There is a suggestion, however, that some species use learned signals to label conspecific individuals in their own communication system when mimicking individually distinctive calls. Bottlenose dolphins (Tursiops truncatus) are a promising animal for exploration in this area because they are capable of vocal production learning and can learn to use arbitrary signals to report the presence or absence of objects. Bottlenose dolphins develop their own unique identity signal, the signature whistle. This whistle encodes individual identity independently of voice features. The copying of signature whistles may therefore allow animals to label or address one another. Here, we show that wild bottlenose dolphins respond to hearing a copy of their own signature whistle by calling back. Animals did not respond to whistles that were not their own signature. This study provides compelling evidence that a dolphin's learned identity signal is used as a label when addressing conspecifics. Bottlenose dolphins therefore appear to be unique as nonhuman mammals to use learned signals as individually specific labels for different social companions in their own natural communication system.


Differential orientation of 10T1/2 mesenchymal cells on non-uniform stretch environments.
Non-uniform stress and strain fields are prevalent in many tissues in vivo, and often exacerbated by disease or injury. These mechanical gradients potentially play a role in contributing to pathological conditions, presenting a need for experimental tools to allow investigation of cell behavior within non-uniformly stimulated environments. Herein, we employ two in vitro cell-stretching devices (one previously published; one newly presented) capable of subjecting cells to cyclic, non-uniform stretches upon the surface of either a circular elastomeric membrane or a cylindrical PDMS tube. After 24 hours of cyclic stretch, 10T1/2 cells on both devices showed marked changes in long-axis orientation, with tendencies to align parallel to the direction of minimal deformation. The degree of this response varied depending on location within the stretch gradients. These results demonstrated the feasibility of conducting cell mechanobiology investigations with the two novel devices, while also highlighting the experimental capabilities of non-uniform mechanical environments for these types of studies. Such capabilities include robust data collection for developing mechanobiological dose-response curves, signal threshold identification, and potential spatial targeting for drug delivery.


Gotta Catch 'Em All: Impact of Pokémon Go on Physical Activity, Sitting Time, and Perceptions of Physical Activity and Health at Baseline and Three-Month Follow-Up.
Objective: The objective was to examine differences in physical activity, sitting time and perceptions of physical activity and health between Pokémon Go users and nonusers at baseline (launch of the application in the United Kingdom) and 3-month follow-up. Materials and Methods: The self-administered, short version of the 7-day recall, International Physical Activity Questionnaire, was adapted to develop the "Physical Activity and Pokémon Go questionnaire," which was distributed using social media. Four weeks after the launch of the application, 461 participants (n = 193 male, n = 265 female, n = 3 transgender) had completed the questionnaire. At 3-month follow-up, 127 participants repeated the questionnaire. Results: At baseline, mixed models analysis of variance (ANOVA) revealed main effects for Pokémon Go users versus nonusers in the amount of days of vigorous physical activity, moderate physical activity, and walking (All P < 0.01). Users reported that they undertook less days of vigorous physical activity than nonusers, but more days of moderate physical activity and walking. There were no differences in body mass index, minutes of vigorous or moderate physical activity, and walking, or sitting on weekdays (All P > 0.05). Repeated measures ANOVA identified increased sitting on weekdays (P < 0.05), but maintained vigorous, moderate and walking physical activity behaviors in users who remained users. Conclusion: Pokémon Go use can increase the frequency of days of physical activity benefitting health. Users at both time points maintained their physical activity behaviour, but increased sitting time on weekdays, highlighting that another intervention to prevent sitting is needed.


The changing attitudes of health professionals and consumers towards a coordinated care trial--SA HealthPlus.
The national coordinated care trials have been a vehicle for health reform in Australia, driven by escalating health care costs and projections of an ageing population. The first round of trials conducted between 1997 and 1999 set the trials a challenge to reduce financial and system barriers to enable health professionals in all sectors and consumers to develop service delivery models which would give better outcomes for patients within existing resources. As part of a change management strategy, the developers of the SA HealthPlus trial assessed the attitudes of health professionals and consumers involved in designing the projects which made up the larger trial, prior to trial development and twelve months later. This paper reports on the results of the survey and how initial enthusiasm gave way to appropriate anxiety as the complexities of creating a new system of care from reactive to prospective patient centred care planning, became a reality. The survey enabled trial developers to show evidence of acceptability for the new model of care and identify areas of concern and appropriate strategies for the project teams. This type of survey and the issues identified may be of benefit to the second round coordinated care trials and health regions aiming to initiate coordinated care programs.


Sub-lethal levels of electric current elicit the biosynthesis of plant secondary metabolites.
Many secondary metabolites that are normally undetectable or in low amounts in healthy plant tissue are synthesized in high amounts in response to microbial infection. Various abiotic and biotic agents have been shown to mimic microorganisms and act as elicitors of the synthesis of these plant compounds. In the present study, sub-lethal levels of electric current are shown to elicit the biosynthesis of secondary metabolites in transgenic and non-transgenic plant tissue. The production of the phytoalexin (+)-pisatin by pea was used as the main model system. Non-transgenic pea hairy roots treated with 30-100 mA of electric current produced 13 times higher amounts of (+)-pisatin than did the non-elicited controls. Electrically elicited transgenic pea hairy root cultures blocked at various enzymatic steps in the (+)-pisatin biosynthetic pathway also accumulated intermediates preceding the blocked enzymatic step. Secondary metabolites not usually produced by pea accumulated in some of the transgenic root cultures after electric elicitation due to the diversion of the intermediates into new pathways. The amount of pisatin in the medium bathing the roots of electro-elicited roots of hydroponically cultivated pea plants was 10 times higher 24 h after elicitation than in the medium surrounding the roots of non-elicited control plants, showing not only that the electric current elicited (+)-pisatin biosynthesis but also that the (+)-pisatin was released from the roots. Seedlings, intact roots or cell suspension cultures of fenugreek (Trigonella foenum-graecum), barrel medic, (Medicago truncatula), Arabidopsis thaliana, red clover (Trifolium pratense) and chickpea (Cicer arietinum) also produced increased levels of secondary metabolites in response to electro-elicitation. On the basis of our results, electric current would appear to be a general elicitor of plant secondary metabolites and to have potential for application in both basic and commercial research.


Qualitative situational analysis of palliative care for adolescents with cancer and HIV in South Africa: healthcare worker perceptions.
The unmet palliative care need is intensified in resource-limited countries where there is inequitable access to healthcare and adolescents and young adults (AYA) fall between the cracks of paediatric and adult services. We aimed to examine palliative care for AYA with cancer and/or HIV in South Africa. Individual and small group interviews using process mapping. Data were collected between November 2016 and March 2017 with staff from government hospitals, hospices and clinics from three provinces of South Africa. Health and social care professionals (n=30) (eg, physicians, nurses, social workers, psychologists) from KwaZulu-Natal, Western Cape and Gauteng provinces. In this qualitative situational analysis study, participants were recruited to interviews and process mapping used to try and identify palliative care pathways of AYA with palliative care needs. We developed a thematic framework using a deductive approach to analyse the interview data. Palliative care pathways were not easily identifiable, ambiguous and varied and participants reported on their experiences and perceptions of service provision. Participants expressed that resource challenges limited the service provision. A need for nurse prescribing of pain relief at community level was mentioned as AYA are reluctant to use hospitals and clinics. Lack of (1) palliative care education among professionals, (2) public awareness and (3) adolescent-specific care posed a challenge to service provision. The changing burden of HIV and increase in cancer prevalence calls for integration of palliative care in mainstream health services. Despite good policies, the pragmatic aspect of training professionals will influence a change in practice. The main task will be to absorb palliative care philosophy and practice at all levels of care to avoid overburdening the care system and to integrate it in community care programmes.


Infratentorial atrophy on magnetic resonance imaging and disability in multiple sclerosis.
Loss of tissue volume in the central nervous system may provide an index of fixed neurological dysfunction in multiple sclerosis. Recent magnetic resonance studies have shown a modest relationship between clinical disability rating scores and transverse sectional area of the cervical spinal cord. To explore further the relationship between atrophy and disability in multiple sclerosis, we estimated the volumes of infratentorial structures from MRIs in a cross-sectional study of 41 patients, 21 with relapsing-remitting multiple sclerosis and 20 with secondary progressive multiple sclerosis. We used the Cavalieri method of modern design stereology with point counting to estimate the volume of brainstem, cerebellum and upper cervical spinal cord from three-dimensional MRIs acquired with an MPRAGE (Magnetization-prepared Rapid Acquisition Gradient Echo) sequence. The volume of the upper (C1-C3) cervical spinal cord was significantly correlated with a composite spinal cord score derived from the appropriate Functional Scale scores of the Expanded Disability Status Scale (r = -0.50, P < 0.01). The cerebellar (r = 0.49, P < 0.01) and brainstem (r = 0.34, P < 0.05) volumes correlated with the Scripp's Neurological Disability Rating Scale scores. The upper cervical cord volumes (r = -0.39, P < 0.01), but not the brainstem or cerebellar volumes, were significantly associated with disease duration. MRI-estimated structural volumes may provide a simple index of axonal and/or myelin loss, the presumed pathological substrates of irreversible impairment and disability in multiple sclerosis.


Nutrient intake and ovarian cancer: an Italian case-control study.
The role of selected macronutrients, cholesterol, and fatty acids in the etiology of epithelial ovarian cancer was analyzed using data from a case-control study carried out in five Italian areas between January 1992 and December 1999. Cases comprised 1,031 women with incident, histologically confirmed epithelial ovarian cancer, admitted to the major teaching and general hospitals of the study areas. Controls comprised 2,411 women admitted for acute, non-neoplastic conditions to the same network of hospitals. Information on dietary habits was elicited using a validated food-frequency questionnaire including 78 food groups and recipes. Odds ratios (OR) and their corresponding 95% confidence intervals (CI) were computed by subsequent quintiles of nutrient intake. Direct associations with ovarian cancer emerged for starch intake (OR = 1.4 in the highest vs the lowest quintile of intake; 95% CI 1.1-1.8), while inverse associations emerged for monounsaturated (OR=0.7; 95% CI 0.5-0.9), and polyunsaturated (OR = 0.7; 95% CI 0.5-0.9) fatty acids. Among fatty acids, oleic (OR = 0.7; 95% CI 0.5-0.9), linoleic (OR = 0.7; 95% CI 0.5-0.9), and linolenic (OR = 0.8; 95% CI 0.6-1.0) acids were inversely related to ovarian cancer. When, however, six macronutrients were included in the same model, only the adverse effect of high starch intake remained significant. Results were consistent in separate strata of menopausal status, parity, and energy intake. Starch was directly associated, and unsaturated fatty acids were inversely associated, with ovarian cancer risk.


Comorbidities, timing of treatments, and chemotherapy use influence outcomes in stage III colon cancer: A population-based European study.
For stage III colon cancer (CC), surgery followed by chemotherapy is the main curative approach, although optimum times between diagnosis and surgery, and surgery and chemotherapy, have not been established. We analysed a population-based sample of 1912 stage III CC cases diagnosed in eight European countries in 2009-2013 aiming to estimate: (i) odds of receiving postoperative chemotherapy, overall and within eight weeks of surgery; (ii) risks of death/relapse, according to treatment, Charlson Comorbidity Index, time from diagnosis to surgery for emergency and elective cases, and time from surgery to chemotherapy; and (iii) time-trends in chemotherapy use. Overall, 97% of cases received surgery and 65% postoperative chemotherapy, with 71% of these receiving chemotherapy within eight weeks of surgery. Risks of death and relapse were higher for cases starting chemotherapy with delay, but better than for cases not given chemotherapy. Fewer patients with high comorbidities received chemotherapy than those with low (P < 0.001). Chemotherapy timing did not vary (P = 0.250) between high and low comorbidity cases. Electively-operated cases with low comorbidities received surgery more promptly than high comorbidity cases. Risks of death and relapse were lower for elective cases given surgery after four weeks than cases given surgery within a week. High comorbidities were always independently associated with poorer outcomes. Chemotherapy use increased over time. Our data indicate that promptly-administered postoperative chemotherapy maximizes its benefit, and that careful assessment of comorbidities is important before treatment. The survival benefit associated with slightly delayed elective surgery deserves further investigation.


Guamanian neurodegenerative disease: are diabetes mellitus and altered humoral immunity clues to pathogenesis?
On the western Pacific island of Guam, parkinsonism, dementia, and amyotrophic lateral sclerosis are highly prevalent but the cause is not known. To assess the possibility that the pathologic process extends beyond the nervous system, we studied patients with Guamanian neurodegenerative disease (N = 16) and Guamanian Chamorro control subjects (N = 16) in the Clinical Research Center of the Mayo Clinic, Rochester, MN. The principal abnormalities found in those with neurodegenerative disease included diabetes mellitus in 44%, elevated levels of serum immunoglobulin A (IgA) in 50%, and elevated IgG in 44%. The mean serum IgM level in the patient group was significantly lower than in the control group. Diabetes mellitus and elevated IgA and IgG levels were also present in 31% of neurologically normal Guamanian subjects. Some of these control subjects, however, probably have preclinical neurodegenerative disease, as found in previously published postmortem studies. Extensive serologic testing did not reveal any consistent profile of autoimmunity. Other blood and urine studies failed to identify hematologic, nutritional, renal, hepatic, or metabolic abnormalities that distinguished patients. Whether diabetes mellitus or abnormalities of immune regulation share common etiopathology with Guamanian neurodegenerative disease deserves further study.


Development and evolution of the collopallium in amniotes: a new hypothesis of field homology.
Embryological development is uniquely positioned to illuminate both hodology in adult brains and its inherited genetic bases. The lateral corner of the lateral ventricle in mammals is a particularly crucial region where cell migration patterns, transiently formed connections, axonal growth kinetics, and fasciculation patterns are complex and interactive. Based on hodology, the sauropsid anterior dorsal ventricular ridge (ADVR) has been proposed as the homologue on a one-to-one basis of the mammalian lateral neocortex (LNC), the basolateral amygdalar complex (BLA), or the claustrum-endopiriform nucleus (CE). Data on gene expression patterns during development have indicated ADVR homology with parts of the latter two structures rather than with LNC. Collothalamic nuclei (the set of dorsal thalamic nuclei that receive their predominant input from the midbrain roof) project to part of BLA and to LNC. Recent findings demonstrate a complex pattern of mutually overlapping but noncongruent gene expression territories and collothalamic projections, which suggests a new, collopallial field hypothesis that the ADVR is homologous as a field to all three structures LNC, BLA, and CE. This hypothesis accounts for current hodological and developmental data as well as for lack of a CE in monotremes and for an abnormal subcortical lamina of gray matter that results from a genetic abnormality in humans.


Drug therapy in the management of acute renal failure.
Pharmacologic intervention aimed at altering the natural history of acute renal failure is a routine practice without scientific support of efficacy. Oliguria has become a separate disease entity with an apparent disregard for the underlying condition that caused it. Volume expansion is clearly beneficial in preventing many volume depleted patients from progressing to acute renal failure. While mannitol and furosemide have been used to "convert" oliguric acute renal failure to the more easy to manage non-oliguric acute renal failure, published reports suggest that responders were not as ill as non-responders. The use of dopamine to increase urine flow in patients with established acute renal failure is the current fashion, but there is little evidence that this drug raises the glomerular filtration rate or shortens the course of acute renal failure. These pharmacologic therapies increase the complexity and cost of care with little tangible evidence of benefit to the patient or the physician caring for the patient.


Nonparaxial propagation of the chirped Airy vortex beams in uniaxial crystal orthogonal to the optical axis.
We study both analytically and numerically nonparaxial propagation dynamics of the Chirped Airy vortex (CAiV) beams in uniaxial crystal orthogonal to the optical axis. The propagation trajectory, the intensity, the radiation forces, the Poynting vector and the angular momentum (AM) of the CAiV beams are illustrated by numerical examples. The influences of the ratio of the extraordinary refractive index to the ordinary refractive index, the linear chirp factor and the quadratic chirp factor on the nonparaxial evolution of the CAiV beams are examined in detail. Results show that the linear chirp factor provides an intensity concentration, which is totally different with the influence of the quadratic chirp. Besides, the uniaxial crystals with different refractive index ratios can be used to control the intensity of optical lobes. Moreover, the value and the direction of the radiation forces depend on the propagation distance and the chirp factor. The chirp factor acting on the Poynting vector and the AM mainly occurs in the direction of vectors. The nonparaxial propagation characteristics of the CAiV beams provide a convenient method to the intensity modulation and the optical manipulation of micro particles.


Elucidating the Elementary Reaction Pathways and Kinetics of Hydroxyl Radical-Induced Acetone Degradation in Aqueous Phase Advanced Oxidation Processes.
Advanced oxidation processes (AOPs) that produce highly reactive hydroxyl radicals are promising methods to destroy aqueous organic contaminants. Hydroxyl radicals react rapidly and nonselectively with organic contaminants and degrade them into intermediates and transformation byproducts. Past studies have indicated that peroxyl radical reactions are responsible for the formation of many intermediate radicals and transformation byproducts. However, complex peroxyl radical reactions that produce identical transformation products make it difficult to experimentally study the elementary reaction pathways and kinetics. In this study, we used ab initio quantum mechanical calculations to identify the thermodynamically preferable elementary reaction pathways of hydroxyl radical-induced acetone degradation by calculating the free energies of the reaction and predicting the corresponding reaction rate constants by calculating the free energies of activation. In addition, we solved the ordinary differential equations for each species participating in the elementary reactions to predict the concentration profiles for acetone and its transformation byproducts in an aqueous phase UV/hydrogen peroxide AOP. Our ab initio quantum mechanical calculations found an insignificant contribution of Russell reaction mechanisms of peroxyl radicals, but significant involvement of HO2• in the peroxyl radical reactions. The predicted concentration profiles were compared with experiments in the literature, validating our elementary reaction-based kinetic model.


Inflammatory pseudotumor of the choroid plexus.
A case of inflammatory pseudotumor is described, in which CT revealed a hyperdense intraventricular lesion with a trapped temporal horn, and MR imaging showed decreased T1 and markedly hypointense T2 signal with homogeneously intense enhancement. Radiologically and grossly the tumor resembled a meningioma; however, histologically it was composed purely of inflammatory, nonneoplastic components. The lesion was resected with no evidence of recurrence at 5-month follow-up.


Effect of alternative antibiotics in treatment of cefotaxime resistant spontaneous bacterial peritonitis.
To evaluate effective alternative antibiotics in treatment of cefotaxime-resistant spontaneous bacterial peritonitis. One hundred cirrhotic patients with spontaneous bacterial peritonitis [ascitic fluid polymorphonuclear cell count (PMNLs) ≥ 250 cells/mm(3) at admission] were empirically treated with cefotaxime sodium 2 g/12 h and volume expansion by intravenous human albumin. All patients were subjected to history taking, complete examination, laboratory tests (including a complete blood cell count, prothrombin time, biochemical tests of liver and kidney function, and fresh urine sediment), chest X-ray, a diagnostic abdominal paracentesis, and the sample subjected to total and differential cell count, chemical examination, aerobic and anaerobic cultures. Patients were divided after 2 d by a second ascitic PMNL count into group I; patients sensitive to cefotaxime (n = 81), group II (n = 19); cases resistant to cefotaxime (less than 25% decrease in ascitic PMNL count). Patients of group II were randomly assigned into meropenem (n = 11) or levofloxacin (n = 8) subgroups. All patients performed an end of treatment ascitic PMNL count. Patients were considered improved when: PMNLs decreased to < 250 cells/mm(3), no growth in previously positive culture cases, and improved clinical manifestations with at least 5 d of antibiotic therapy. Age, sex, and Child classes showed no significant difference between group I and group II. Fever and abdominal pain were the most frequent manifestations and were reported in 82.7% and 80.2% of patients in group I and in 94.7% and 84.2% of patients in group II, respectively. Patients in group II had a more severe ascitic inflammatory response than group I and this was demonstrated by more ascitic lactate dehydrogenase (LDH) [median: 540 IU/L (range: 150-1200 IU/L) vs median: 240 IU/L (range: 180-500 IU/L), P = 0.000] and PMNL [median: 15,000 cell/mm(3) (range: 957-23,822 cell/mm(3)) vs 3400 cell/mm(3) (range: 695-26,400 cell/mm(3)), P = 0.000] counts. Ascitic fluid culture was positive in 32% of cases. Cefotaxime failed in 19% of patients; of these patients, 11 (100%) responded to meropenem and 6 (75%) responded to levofloxacin. Two patients with failed levofloxacin therapy were treated according to the in vitro culture and sensitivity (one case was treated with vancomycin and one case was treated with ampicillin/sulbactam). In group II the meropenem subgroup had higher LDH (range: 108-860 IU/L vs 120-491 IU/L, P = 0.042) and PMNL counts (range: 957-23,822 cell/mm(3)vs 957-15,222 cell/mm(3), P = 0.000) at initiation of the alternative antibiotic therapy; there was no significant difference in the studied parameters between patients responsive to meropenem and patients responsive to levofloxacin at the end of therapy (mean ± SD: 316.01 ± 104.03 PMNLs/mm(3)vs 265.63 ± 69.61 PMNLs/mm(3), P = 0.307). The isolated organisms found in group II were; enterococci, acinetobacter, expanded-spectrum β-lactamase producing Escherichia coli, β-lactamase producing Enterobacter and Staphylococcus aureus. Empirical treatment with cefotaxime is effective in 81% of cases; meropenem is effective in cefotaxime-resistant cases.


Time to Treatment Intensification After Monotherapy Failure and Its Association With Subsequent Glycemic Control Among 93,515 Patients With Type 2 Diabetes.
The goal of this study was to evaluate the association between the timing of treatment intensification and subsequent glycemic control among patients with type 2 diabetes in whom monotherapy fails. This retrospective analysis of the U.K. Clinical Practice Research Datalink database focused on patients with type 2 diabetes and one or more HbA1c measurements ≥7% (≥53 mmol/mol) after ≥3 months of metformin or sulfonylurea monotherapy (first measurement meeting these criteria was taken as the study index date). Baseline (6 months before the index date) characteristics were stratified by time from the index date to intensification (early: <12 months; intermediate: 12 to <24 months; late: 24 to <36 months). Intensification was defined as initiating after the index date one or more noninsulin antidiabetes medication in addition to metformin or a sulfonylurea. Association between time to intensification and subsequent glycemic control (first HbA1c <7% [<53 mmol/mol] after intensification) was evaluated using Kaplan-Meier analyses and Cox proportional hazard models that accounted for baseline differences. Of the 93,515 patients who met the study criteria (mean age 60 years; ∼59% male; 80% taking metformin), 23,761 (25%) intensified <12 months after the index date; 11,908 (13%) intensified after 12 to <24 months; and 7,146 (8%) intensified after 24 to <36 months. Patients who intensified treatment ≥36 months after the index date (n = 9,638 [10%]) and those with no evidence of treatment intensification during the observable follow-up period (n = 41,062 [44%]) were not included in further analyses. The median times from intensification to control were 20.0, 24.1, and 25.7 months, respectively, for the early, intermediate, and late intensification cohorts. After adjustment for baseline differences, the likelihood of attaining glycemic control was 22% and 28% lower for patients in the intermediate and late intensification groups, respectively, compared with those intensifying early (P < 0.0001). Earlier treatment intensification is associated with shorter time to subsequent glycemic control, independent of whether patients initiate first-line treatment with metformin or a sulfonylurea.


Estimating propensity scores and causal survival functions using prevalent survival data.
This article develops semiparametric approaches for estimation of propensity scores and causal survival functions from prevalent survival data. The analytical problem arises when the prevalent sampling is adopted for collecting failure times and, as a result, the covariates are incompletely observed due to their association with failure time. The proposed procedure for estimating propensity scores shares interesting features similar to the likelihood formulation in case-control study, but in our case it requires additional consideration in the intercept term. The result shows that the corrected propensity scores in logistic regression setting can be obtained through standard estimation procedure with specific adjustments on the intercept term. For causal estimation, two different types of missing sources are encountered in our model: one can be explained by potential outcome framework; the other is caused by the prevalent sampling scheme. Statistical analysis without adjusting bias from both sources of missingness will lead to biased results in causal inference. The proposed methods were partly motivated by and applied to the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked data for women diagnosed with breast cancer.


Coping with Dravet syndrome: parental experiences with a catastrophic epilepsy.
The aim of this study was to investigate how parents cope with and care for a child with Dravet syndrome, a severe myoclonic epilepsy with three distinct stages. Twenty-four parents of children with Dravet syndrome participated in a semi-structured interview and 17 completed the Impact of Childhood Neurologic Disability Scale (ICND) questionnaire. Children included 11 males and 13 females aged 2 to 24 years (mean age 10y 2mo [SD 5y 8mo]). Stage 1 of the syndrome was generally very difficult. Uncertainty about the diagnosis was the primary cause of stress. Seizure control was worst during this time. The primary concern in Stage 2 remained seizure control but developmental, behavioral, and sleep issues also emerged. Negative effects were noted in parents' relationships with others. Stage 3 brought better seizure control but decreased cognitive level, increased behavioral problems, and increasing social isolation for parents. Respite and relief care were hard to obtain at each stage. ICND scores mirrored the findings of the interviews. The three stages of Dravet syndrome present serious challenges for parents. Seizures are persistent and severe but developmental, behavioral, and sleep issues add to the stress. In the absence of successful medical treatment for Dravet syndrome, further attention needs to be paid to helping families cope with the disorder.


The use of a milli-whistle as a detector in gas analysis by gas chromatography.
This mini-review introduces a general understanding of the use of a milli-whistle as a gas chromatography (GC) detector in gas analysis, including our research on the methodology and theory associated with a number of different related applications. The milli-whistle is connected to the outlet of a GC capillary, and when the eluted gases and the GC carrier gas pass through it, a sound with a fundamental frequency is produced. The sound wave can be picked up by a microphone or an accelerometer, and after a fast Fourier transform, the online data obtained for frequency-change vs. retention time constitute a new method for detecting gases. The first part of this review discusses the fundamentals of the milli-whistle. Some modifications are also discussed, including various types of whistles and an attempt to maximize the sensitivity and stability of the method. The second part then focuses on several practical applications, including an analysis of hydrogen released from ammonia borane, inorganic gases produced from fireworks, the CO2/O2 ratio from expired human breath and a purity test for alcohols. These studies show that the GC-whistle method has great potential for use as a fast sampling ionization method, and for the direct analysis of biological and chemical samples at under ambient conditions.


Cortical Gyrification Morphology in ASD and ADHD: Implication for Further Similarities or Disorder-Specific Features?
Shared etiological pathways are suggested in ASD and ADHD given high rates of comorbidity, phenotypic overlap and shared genetic susceptibility. Given the peak of cortical gyrification expansion and emergence of ASD and ADHD symptomology in early development, we investigated gyrification morphology in 539 children and adolescents (6-17 years of age) with ASD (n=197) and ADHD (n=96) compared to typically developing controls (n=246) using the local Gyrification Index (lGI) to provide insight into contributing etiopathological factors in these two disorders. We also examined IQ effects and functional implications of gyrification by exploring the relation between lGI and ASD and ADHD symptomatology beyond diagnosis. General Linear Models yielded no group differences in lGI, and across groups, we identified an age-related decrease of lGI and greater lGI in females compared to males. No diagnosis-by-age interactions were found. Accounting for IQ variability in the model (n=484) yielded similar results. No significant associations were found between lGI and social communication deficits, repetitive and restricted behaviours, inattention or adaptive functioning. By examining both disorders and controls using shared methodology, we found no evidence of atypicality in gyrification as measured by the lGI in these conditions.


Impaired neurite development associated with mitochondrial dysfunction in dopaminergic neurons differentiated from exfoliated deciduous tooth-derived pulp stem cells of children with autism spectrum disorder.
Autism spectrum disorder (ASD) is a highly heterogeneous neurodevelopmental disorder characterized by impaired social interactions, restrictive interests, and repetitive stereotypic behaviors. Among the various mechanisms underlying the pathogenesis of ASD, dysfunctions of dopaminergic signaling and mitochondria have been hypothesized to explain the core symptoms of children with ASD. However, only a few studies focusing on the pathological association between dopaminergic neurons (DN) and mitochondria in ASD have been performed using patient-derived stem cells and in vitro differentiated neurons. Stem cells from human exfoliated deciduous teeth (SHED) are neural crest-derived mesenchymal stem cells present in the dental pulp of exfoliated deciduous teeth; these cells can differentiate into dopaminergic neurons (DN) in vitro. This study aimed to investigate the pathological association between development of DN and mitochondria in ASD by using SHED as a disease- or patient-specific cellular model. The SHED obtained from three children with ASD and three typically developing children were differentiated into DN, and the neurobiology of these cells was examined. The DN derived from children with ASD showed impaired neurite outgrowth and branching, associated with decreased mitochondrial membrane potential, ATP production, number of mitochondria within the neurites, amount of mitochondria per cell area and intracellular calcium level. In addition, impaired neurite outgrowth and branching of ASD-derived DN were not improved by brain-derived neurotrophic factor (BDNF), suggesting impairment of the BDNF signaling pathway in ASD. These results imply that intracerebral dopamine production may have decreased in these children. The earliest age at which deciduous teeth spontaneously exfoliate in humans, and SHED can be noninvasively collected, is approximately 6 years. Our results suggest that in vitro analysis of SHED-derived DN obtained from children with ASD provides neurobiological information that may be useful in determining treatment strategies in the early stages of ASD.


Multiparametric molecular imaging provides mechanistic insights into sympathetic innervation impairment in the viable infarct border zone.
Impaired catecholamine handling in the viable infarct border zone may play an important role in ventricular remodeling and lethal arrhythmia. We sought to get further biologic insights into cardiac sympathetic neuronal pathology after myocardial infarction, using multiple tomographic imaging techniques. In a porcine model of myocardial infarction (n = 13), PET and MR imaging were performed after 4-6 wk and integrated with electrophysiologic testing and postmortem histology. PET with the physiologic neurotransmitter (11)C-epinephrine, which is sensitive to metabolic degradation unless it is stored and protected in neuronal vesicles, identified a defect exceeding the perfusion defect (defined by (13)N-ammonia; defect size in all animals, 42 ± 12 vs. 35% ± 12% of left ventricle, P < 0.001). In a subgroup of 7 animals, defect of the metabolically resistant catecholamine (11)C-hydroxyephedrine was smaller than epinephrine (41 ± 8 vs. 47% ± 6% of left ventricle, P = 0.004), whereas defect of a third catecholamine, (11)C-phenylephrine, which is sensitive to metabolic degradation, was similar to epinephrine (48 ± 6 vs. 47% ± 6%, P = 0.011 vs. perfusion defect). Histology confirmed the presence of nerve fibers in the infarct border zone. Tagged MR imaging identified impaired peak circumferential wall strain and wall thickening in myocardial segments with epinephrine/perfusion mismatch (n = 6). Confirmatory of prior work, inducible ventricular tachycardia was associated with a larger epinephrine/perfusion mismatch (n = 11). In the viable infarct border zone, neuronal vesicular catecholamine storage and protection from metabolic degradation are more severely altered than catecholamine uptake. This alteration may reflect an intermediate state between normal innervation and complete denervation in advanced disease.


The genetic landscape of clinical resistance to RAF inhibition in metastatic melanoma.
Most patients with BRAF(V600)-mutant metastatic melanoma develop resistance to selective RAF kinase inhibitors. The spectrum of clinical genetic resistance mechanisms to RAF inhibitors and options for salvage therapy are incompletely understood. We performed whole-exome sequencing on formalin-fixed, paraffin-embedded tumors from 45 patients with BRAF(V600)-mutant metastatic melanoma who received vemurafenib or dabrafenib monotherapy. Genetic alterations in known or putative RAF inhibitor resistance genes were observed in 23 of 45 patients (51%). Besides previously characterized alterations, we discovered a "long tail" of new mitogen-activated protein kinase (MAPK) pathway alterations (MAP2K2, MITF) that confer RAF inhibitor resistance. In three cases, multiple resistance gene alterations were observed within the same tumor biopsy. Overall, RAF inhibitor therapy leads to diverse clinical genetic resistance mechanisms, mostly involving MAPK pathway reactivation. Novel therapeutic combinations may be needed to achieve durable clinical control of BRAF(V600)-mutant melanoma. Integrating clinical genomics with preclinical screens may model subsequent resistance studies.


Evidence of the Red-Queen Hypothesis from Accelerated Rates of Evolution of Genes Involved in Biotic Interactions in Pneumocystis.
Pneumocystis species are ascomycete fungi adapted to live inside the lungs of mammals. These ascomycetes show extensive stenoxenism, meaning that each species of Pneumocystis infects a single species of host. Here, we study the effect exerted by natural selection on gene evolution in the genomes of three Pneumocystis species. We show that genes involved in host interaction evolve under positive selection. In the first place, we found strong evidence of episodic diversifying selection in Major surface glycoproteins (Msg). These proteins are located on the surface of Pneumocystis and are used for host attachment and probably for immune system evasion. Consistent with their function as antigens, most sites under diversifying selection in Msg code for residues with large relative surface accessibility areas. We also found evidence of positive selection in part of the cell machinery used to export Msg to the cell surface. Specifically, we found that genes participating in glycosylphosphatidylinositol (GPI) biosynthesis show an increased rate of nonsynonymous substitutions (dN) versus synonymous substitutions (dS). GPI is a molecule synthesized in the endoplasmic reticulum that is used to anchor proteins to membranes. We interpret the aforementioned findings as evidence of selective pressure exerted by the host immune system on Pneumocystis species, shaping the evolution of Msg and several proteins involved in GPI biosynthesis. We suggest that genome evolution in Pneumocystis is well described by the Red-Queen hypothesis whereby genes relevant for biotic interactions show accelerated rates of evolution.


On the question of equilibrium in mercury + thallium iodide arc discharges.
Two aspects of the concept of local thermodynamic equilibrium are examined in mediumpressure, mercury arc discharges with thallium iodide additives. It is shown theoretically that the experimental intensity of the T 5350 A line is reasonably consistent with a Boltzmann distribution of excited states. In fact, it is theoretically and experimentally shown that self-absorption is a major factor in limiting the intensity of the bright thallium 5350 A line even though Ti is a minor constituent in the discharge and even though this line does not terminate on the ground state. The dramatic spectral inhomogeneity that develops in long, horizontal, ac discharges with excess TII in the presence of a longitudinal temperature gradient is examined in detail. Both emission and absorption studies show that the spectral inhomogeneity is caused by a large gradient in the thallium concentration. This inhomogeneous distribution of TlI vapors is caused by an inhomogeneous and unsymmetrical distribution of condensed THI along the tube walls which results from the temperature gradient. In turn, the temperature gradient is increased by the ionization of thallium which locally decreases the electrical resistivity, electric field, and power input to the arc. Thus, an equilibrium distribution of TlI vapors is attained only if the distribution of condensed TlI is reasonably uniform and the tube geometry and pressure are favorable for rapid diffusion of the vapors. The inhomogeneity also disappears if a sufficiently small quantity of solid TII is added to the lamp so that it is all vaporized.


Dysfunction of the retinal pigment epithelium with age: increased iron decreases phagocytosis and lysosomal activity.
Iron accumulation with age in the retinal pigment epithelium (RPE) may be one important source of oxidative stress that contributes to age-related macular degeneration (AMD). Young and old rodent RPE/choroid were compared to assess iron homeostasis during normal aging and the effects of increased iron on the functions of retinal pigment epithelial cells. The iron level, mRNA expression, and protein level of iron-regulatory molecules in RPE/choroid were quantitatively compared between young and old animals. To test the effects of increased intracellular iron on the functions of retinal pigment epithelial cells, in vitro ARPE-19 cells were treated with high levels of iron and assessed for phagocytosis activity and lysosomal activity. Iron level was significantly increased in the aged RPE/choroid. Ferritin and ceruloplasmin mRNAs were significantly increased in the aged RPE/choroid, whereas transferrin, transferrin receptor, and ferroportin mRNAs did not change with age. At the protein level, decreased transferrin and transferrin receptor, increased ferritin and ceruloplasmin, and unchanged ferroportin were observed in the aged RPE/choroid. Exposure of ARPE-19 cells to increased iron markedly decreased phagocytosis activity, interrupted cathepsin D processing, and reduced cathepsin D activity in retinal pigment epithelial cells. The RPE/choroid of aged animals demonstrates iron accumulation and associated alterations in iron homeostasis. Iron accumulation with age may impair the phagocytosis and lysosomal functions of retinal pigment epithelial cells in the aged RPE/choroid. Therefore, age-related changes of iron homeostasis in the RPE could increase the susceptibility of the tissue to genetic mutations associated with AMD.


Enhanced individual trabecular repair and its mechanical implications in parathyroid hormone and alendronate treated rat tibial bone.
Combined parathyroid hormone (PTH) and bisphosphonate (alendronate-ALN) therapy has recently been shown to increase bone volume fraction and plate-like trabecular structure beyond either monotherapy. To identify the mechanism through which plate-like structure was enhanced, we used in vivo microcomputed tomography (μCT) of the proximal tibia metaphysis and individual trabecular dynamics (ITD) analysis to quantify connectivity repair (incidences of rod connection and plate perforation filling) and deterioration (incidences of rod disconnection and plate perforation). Three-month-old female, intact rats were scanned before and after a 12 day treatment period of vehicle (Veh, n = 5), ALN (n = 6), PTH (n = 6), and combined (PTH+ALN, n = 6) therapy. Additionally, we used computational simulation and finite element (FE) analysis to delineate the contributions of connectivity repair or trabecular thickening to trabecular bone stiffness. Our results showed that the combined therapy group had greater connectivity repair (5.8 ± 0.5% connected rods and 2.0 ± 0.3% filled plates) beyond that of the Veh group, resulting in the greatest net gain in connectivity. For all treatment groups, increases in bone volume due to thickening (5-31%) were far greater than those due to connectivity repair (2-3%). Newly formed bone contributing only to trabecular thickening caused a 10%, 41%, and 69% increase in stiffness in the ALN, PTH, and PTH+ALN groups, respectively. Moreover, newly formed bone that led to connectivity repair resulted in an additional improvement in stiffness, with the highest in PTH+ALN (by an additional 12%), which was significantly greater than either PTH (5.6%) or ALN (4.5%). An efficiency ratio was calculated as the mean percent increase in stiffness divided by mean percent increase in BV for either thickening or connectivity repair in each treatment. For all treatments, the efficiency ratio of connectivity repair (ALN: 2.9; PTH: 3.4; PTH+ALN: 4.4) was higher than that due to thickening (ALN: 2.0; PTH: 1.7; PTH+ALN: 2.2), suggesting connectivity repair required less new bone formation to induce larger gains in stiffness. We conclude that through rod connection and plate perforation filling PTH+ALN combination therapy improved bone stiffness in a more efficient and effective manner than either monotherapy.